Jagged2 CRISPR Activation Plasmid (h): sc-401322-ACT

Human JAG2 encodes Jagged2, a membrane-tethered ligand for NOTCH receptors that mediates juxtacrine signaling to regulate cell fate decisions, lineage commitment, and tissue patterning. Jagged2-driven Notch activation influences transcriptional programs controlling proliferation, differentiation, and apoptosis, with strong connections to stem/progenitor maintenance and developmental processes. Dysregulated JAG2/Notch signaling has been associated with altered epithelial and hematopoietic differentiation states and has been investigated in contexts such as oncogenic signaling, immune modulation, and developmental disorders. As a pathway node, Jagged2 is frequently used to probe Notch-dependent crosstalk with Wnt, TGF-β, and inflammatory signaling networks.

Jagged2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous JAG2 expression without altering the underlying DNA sequence.

Jagged2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the JAG2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the JAG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Jagged2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native JAG2 locus and enabling the study of Jagged2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Jagged2 pathway restoration in tumor cells with silenced or reduced JAG2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.